A new technique of high-efficiency, general-purpose natural productsynthesis of gene clusters for multi-point chromosome insertion.

On December 5, 2018, the international academic journal BlycomA Ging published a paper entitled aMSGE: advanced multiplex site-specific genome engineering with orthogonal modular rereses in actinomyctes.
the study has created a new technique for the insertion of multi-point chromosomes in the synthesis of multi-point chromosomes of highly efficient and versatile natural products of line-lending bacteria, providing another advanced operational tool for the research and development of microbial drugs.
natural products of line-and-line bacteria have diversified chemical structure and broad-spectrum biological activity, has been widely used as insecticides, anti-tumor drugs and immunosuppressants, for human health and agricultural pest control has made a great contribution.
However, the bacteria produced by important drugs of line bacteria are usually low in fermentation units and difficult to operate genetically.
at present, the construction of high-yield bacteria mainly relies on conventional mutagenic screening, low efficiency and heavy workload, so it is urgent to develop advanced and efficient molecular breeding platform, so as to achieve the upgrading of its biological manufacturing technology.
to optimize the biosynthesis of the target product, the metabolic engineering strategies used in the current widely used include strengthening the synthesis path, cutting out the competitive branch, optimizing the regulatory network, chambering of the reaction zone and real-time dynamic response.
it self-evident that it is one of the most direct and effective strategies to strengthen target product synthesis by increasing the number of copies of biosynthetic gene clusters.
2017, Jiang Weihong's research team established a new gene cluster amplification method for the synthesis of natural products of line-lending bacteria, based on the concept of "one integrated enzyme - multiple attB sites" (Figure 1A left).
the method significantly accelerates the construction of heterogeneous expression chassis cells and some easy-to-operate industrial strains, however, it takes multiple rounds to introduce artificial attB sites and is not suitable for genetically difficult line-out bacteria.
in this paper, the author cleverly uses multiple attB sites and a series of orthogonal site-specific recombination systems to develop a more efficient and universal gene cluster amplification method aMSGE (Figure 1A right) based on the concept of "multiple integrated enzymes - multiple attB sites".
the new method can be inserted in one step by adding multiple integrated systems directly to vectors containing target gene clusters.
has also built a "plug-and-play" multi-copy integration toolbox (Figure 1B) to speed up the implementation of the aMSGE approach.
finally, using the newly established aMSGE method, the authors quickly realized the chromosomal integration of the a synthetic gene of a propylene dicolcoenzyme and 5-ketamine (pesticide Milbemycin precursor) in the sky blue streptomycin and ice city streptomycin, respectively, and the final product of the end product of the end-product of the growth of reformer in reformerin and 5-ketone-belmycin, respectively, 4.6 and 1.9 times, reflecting the universality of the method.
as a new generation of line-to-line bacteria synthesis biotechnology, the aMSGE method does not require any genetic modification of chromosomes before the target gene (cluster) amplification, thus saving significantly molecular breeding time and is suitable for industrial strains with genetic difficulties.
the aMSGE approach can also be extended to other bacteria's genome engineering to accelerate the efficient synthesis of chemicals, biofuels and drugs, as site-specific recombination systems are widely distributed across a wide range of bacteria. Li Lei, a postdoctoral
, is the first author of the paper and Lu Yinhua, a researcher at Shanghai Normal University, co-author of the paper.
the research was supported by major new drug creation science and technology projects, post-doctoral innovation talent support programs, Shanghai Natural Science Foundation, etc.
Source: Institute of Plant Physiology and Ecology.